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首页> 外文期刊>Journal of materials science >Trans-differentiation of human adipose-derived mesenchymal stem cells into cardiomyocyte-like cells on decellularized bovine myocardial extracellular matrix-based films
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Trans-differentiation of human adipose-derived mesenchymal stem cells into cardiomyocyte-like cells on decellularized bovine myocardial extracellular matrix-based films

机译：人脂肪来源的间充质干细胞在脱细胞牛心肌细胞外基质基膜上的分化为心肌样细胞

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In this study, we aimed at fabricating decellularized bovine myocardial extracellular matrix-based films (dMEbF) for cardiac tissue engineering (CTE). The decellularization process was carried out utilizing four consecutive stages including hypotonic treatment, detergent treatment, enzymatic digestion and decontamination, respectively. In order to fabricate the dMEbF, dBM were digested with pepsin and gelation process was conducted. dMEbF were then crosslinked with N-hydroxysuccinimide/1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide (NHS/EDC) to increase their durability. Nuclear contents of native BM and decellularized BM (dBM) tissues were determined with DNA content analysis and agarose-gel electrophoresis. Cell viability on dMEbF for 3rd, 7th, and 14th days was assessed by MTT assay. Cell attachment on dMEbF was also studied by scanning electron microscopy. Trans-differentiation capacity of human adipose-derived mesenchymal stem cells (hAMSCs) into cardiomyocyte-like cells on dMEbF were also evaluated by histochemical and immunohistochemical analyses. DNA contents for native and dBM were, respectively, found as 886.11 +/- 164.85 and 47.66 +/- 0.09 ng/mg dry weight, indicating a successful decellularization process. The results of glycosaminoglycan and hydroxyproline assay, and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), performed in order to characterize the extracellular matrix (ECM) composition of native and dBM tissue, showed that the BM matrix was not damaged during the proposed method. Lastly, regarding the histological study, dMEbF not only mimics native ECM, but also induces the stem cells into cardiomyocyte-like cells phenotype which brings it the potential of use in CTE.
机译：在这项研究中，我们旨在制造用于心脏组织工程（CTE）的脱细胞牛心肌细胞外基质基膜（dMEbF）。脱细胞过程利用四个连续的阶段进行，分别包括低渗处理，去污剂处理，酶消化和去污。为了制造dMEbF，用胃蛋白酶消化dMb，并进行凝胶化过程。然后将dMEbF与N-羟基琥珀酰亚胺/ 1-乙基-3-（3-二甲基氨基丙基）-碳二亚胺（NHS / EDC）交联以增加其耐久性。通过DNA含量分析和琼脂糖凝胶电泳确定天然BM和脱细胞BM（dBM）组织的核含量。通过MTT分析评估在dMEbF上第3、7和14天的细胞生存力。还通过扫描电子显微镜研究了dMEbF上的细胞附着。还通过组织化学和免疫组织化学分析评估了人脂肪来源的间充质干细胞（hAMSC）在dMEbF上向心肌样细胞的转分化能力。天然和dBM的DNA含量分别为886.11 +/- 164.85和47.66 +/- 0.09 ng / mg干重，表明脱细胞过程成功。为了表征天然和dBM组织的细胞外基质（ECM）成分，进行了糖胺聚糖和羟脯氨酸测定以及十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）的结果，表明在提出的过程中BM基质未受损方法。最后，关于组织学研究，dMEbF不仅模仿天然ECM，而且还诱导干细胞成为心肌样细胞表型，这使其具有用于CTE的潜力。

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    《Journal of materials science》 |2018年第8期|127.1-127.13|共13页
  • 作者

    Arslan Yavuz Emre; Galata Yusuf Furkan; Arslan Tugba Sezgin; Derkus Burak;

  • 作者单位

    Canakkale Onsekiz Mart Univ, Regenerat Biomat Lab, Engn Fac, Dept Bioengn, TR-17100 Canakkale, Turkey;

    Canakkale Onsekiz Mart Univ, Regenerat Biomat Lab, Engn Fac, Dept Bioengn, TR-17100 Canakkale, Turkey;

    Canakkale Onsekiz Mart Univ, Regenerat Biomat Lab, Engn Fac, Dept Bioengn, TR-17100 Canakkale, Turkey;

    Eskisehir Osmangazi Univ, Dept Biomed Engn, Engn Fac, TR-26480 Eskisehir, Turkey;

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